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J. Biol. Chem., Vol. 276, Issue 17, 14443-14450, April 27, 2001

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Erk Is Essential for Growth, Differentiation, Integrin Expression, and Cell Function in Human Osteoblastic Cells^*

Chung-Fang Lai, Lala Chaudhary^§, Aurora Fausto, Linda R. Halstead, Daniel S. Ory^¶, Louis V. Avioli, and Su-Li Cheng

From the  Division of Bone and Mineral Diseases, ^§ Renal Division, and ^¶ Cardiovascular Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110

Extracellular signal-regulated kinases (Erks), members of the mitogen-activated protein kinase superfamily, play an important role in cell proliferation and differentiation. In this study we employed a dominant negative approach to determine the role of Erks in the regulation of human osteoblastic cell function. Human osteoblastic cells were transduced with a pseudotyped retrovirus encoding either a mutated Erk1 protein with a dominant negative action against both Erk1 and Erk2 (Erk1DN cells) or the LacZ protein (LacZ cells) as a control. Both basal and growth factor-stimulated MAPK activity and cell proliferation were inhibited in Erk1DN cells. Expression of Erk1DN protein suppressed both osteoblast differentiation and matrix mineralization by decreasing alkaline phosphatase activity and the deposition of bone matrix proteins. Cell adhesion to collagen, osteopontin, and vitronectin was decreased in Erk1DN cells as compared with LacZ cells. Cell spreading and migration on these matrices were also inhibited. In Erk1DN cells, expression of ₁, _v3, and _v5 integrins on the surface was decreased. Metabolic labeling indicated that the synthesis of these integrins was inhibited in Erk1DN cells. These data suggest that Erks are not only essential for the growth and differentiation of osteoblasts but also are important for osteoblast adhesion, spreading, migration, and integrin expression.

This paper is dedicated to the memory of Dr. Louis V. Avioli.

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